Generation and analysis of cartilage-specific CCN2 overexpression in transgenic mice

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Recent progress in gene-editing technology has provided a strong impact for improved our understanding of molecular functions in living organisms. Here we describe our method to generate transgene-overexpressing mouse models, which method involves the use of tissue-specific promoters for analyzing a certain molecule (s) in special tissues. The protocol described in this chapter uses the Col2a1 promoter-enhancer, which is known for driving specific and strong transgene expression in cartilage and is based on several of our studies showing a positive role of the connective tissue growth factor (CCN2) in cartilage-bone development and maintenance of articular cartilage. These mice show strongly accelerated endochondral ossification resulting in enhanced bone elongation, as well as resistance to age-related articular degeneration. This protocol also describes how to analyze the molecular mechanisms of these phenomena by use of chondrocytes isolated from CCN2-overexpressing cartilage.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages391-403
Number of pages13
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1489
ISSN (Print)1064-3745

Keywords

  • Aging
  • Cartilage
  • Col2a1 promoter-enhancer
  • Development
  • Endochondral ossification
  • Histochemistry
  • IGF
  • LacZ staining
  • Skeletal preparation
  • Transgenic

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Hattori, T., Itoh, S., & Takigawa, M. (2017). Generation and analysis of cartilage-specific CCN2 overexpression in transgenic mice. In Methods in Molecular Biology (pp. 391-403). (Methods in Molecular Biology; Vol. 1489). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6430-7_32